肿瘤坏死因子α在肺炎支原体感染小鼠主要脏器和血清中的表达及其相关机制

doi:10.7694/jldxyxb20130623

吉林大学学报(医学版)

肿瘤坏死因子α在肺炎支原体感染小鼠主要脏器和血清中的表达及其相关机制

宋星宇¹，车广华¹，高航²，胡起波¹，许忠¹，潘薇¹，徐坤³ 

1.吉林大学第二医院儿科，吉林长春 130041；2. 吉林大学白求恩医学院病理学系，吉林长春 130021；3. 吉林大学公共卫生学院营养与食品卫生学教研室，吉林长春 130021

收稿日期:2013-03-25 发布日期:2013-12-12
通讯作者: 潘薇（Tel：0431-88796802，E-mail：pan_wei8874@sina.com） E-mail:pan_wei8874@sina.com
作者简介:宋星宇(1983-),女，吉林省大安市人，医师，医学硕士，主要从事小儿支原体肺炎基础及临床方面的研究。
基金资助:
吉林省科技厅科研基金资助课题（YYZX201123-2）；吉林大学基本科研业务费资助课题（2012）


Expressions of tumor necrosis factor-α in main organs and  serum of mice infected by Mycoplasma pneumoniae and their mechanisms

SONG Xing-yu¹，CHE Guang-hua¹，GAO Hang²，HU Qi-bo¹，XU Zhong¹，PAN Wei¹，XU Kun³

Received:2013-03-25 Published:2013-12-12

摘要/Abstract

摘要：

［摘要］目的：研究肿瘤坏死因子α（TNF-α）在小鼠肺炎支原体（MP）感染模型主要脏器和血清中的表达，阐明其与MP感染的关系。方法：通过PPLO肉汤培养基培养MP，选择72只BABL/C小鼠，随机分为对照组（未感染MP的小鼠）、MP感染组（感染MP的小鼠)和抗生素治疗组(感染MP后应用阿奇霉素治疗的小鼠)，每组24只。鼻腔滴入培养成功的MP，建立MP感染模型。采用ELISA法测定小鼠血清中 TNF-α的水平;提取小鼠肺组织中的细胞总RNA，应用RT-PCR 法检测小鼠肺组织中TNF-α mRNA 的表达水平;采用免疫组织化学技术观察MP感染小鼠肺、肝脏、肾脏和心脏的病变程度。结果：成功建立MP感染小鼠模型，MP菌液进入小鼠呼吸系统后，MP感染组和抗生素治疗组小鼠出现活动减少、进食差、精神萎靡、鼻部分泌物增加、咳嗽、竖毛和寒战等情况，对照组小鼠进食、饮水和活动度无明显改变。ELISA法，MP感染组小鼠血清中TNF-α水平明显高于对照组与抗生素治疗组（P<0.05），抗生素治疗组小鼠血清中TNF-α的水平高于对照组（P<0.05）；RT-PCR法，MP感染组小鼠肺组织中TNF-α mRNA水平高于对照组和抗生素治疗组（P<0.05），且抗生素治疗组小鼠TNF-α　mRNA水平高于对照组（P<0.05）；免疫组织化学法，MP感染组小鼠肺组织病变重于抗生素治疗组和对照组，且抗生素治疗组重于对照组，对照组无病变或轻微病变（P<0.05），3种实验方法表达结果一致。结论：TNF-α阳性表达强度与MP肺炎的病情严重程度有关联。

关键词: 肺炎支原体, 肿瘤坏死因子&alpha, 酶联免疫吸附实验, 免疫组织化学技术

Abstract:

Abstract:Objective To investigate the expressions of tumor necrosis factor-α (TNF-α) in main organs and serum of mouse model infected by Ｍycoplasma pneumonia (MP) and to clarify the relationship between TNF-α and MP infection.Methods MP was cultured in the medium containing PPLO broth.Total 72 BABL/C mice were randomly divided into control group(non MP-infected mice，n=24),MP infection group (MP-infected mice，n=24)and antibiotic treatment group(MP-infected mice treated with azithromycin，n=24).MP infection models were established with the method of dripping cultured MP into nasal cavity of mice.ELISA method was used to assay the levels of TNF-α in the serum of the mice in various groups.Total RNA in the lung tissue of mice was extracted and RT-PCR method was used to detect the expression levels of TNF-α mRNA.Immunohistochemical technique was applied to show the lesion of lung,liver,kidney,and heart tissues in the MP-infected mice.Results The MP-infected mouse models were established successfully,and MP strains were dripped into the respiratory system of mice.The activities of the mice in MP infection group and antibiotic treatment group were reduced,and they showed poor eating,listlessness,nasal secretions,cough,piloerection,chills ect.Contrastly,there were no changes in eating,drinking and activity of the mice in control group.The ELISA results showed that the level of serum TNF-α of the mice in MP infection group was higher than those in antibiotic treatment group and control group(P<0.05）.At the same time,the level of serum TNF-α of the mice in antibiotic treatment group was higher than that in control group(P<0.05）.The RT-PCR results showed the level of TNF-α mRNA in lung tissue of the mice in MP infection group was higher than those in control group and antibiotic treatment group(P<0.05),and the level of TNF-α mRNA of the mice in MP treatment group was higher than that in control group(P<0.05).The imunohistochemical results showed the lesion of lung tissue of the mice in MP infection group was heavier than those in antibiotic treatment group and control group,and it was sligher in antibiotic treatment group than control group,while there was no disease or minor lesion of lung tissue of the mice in control group（P<0.05）. The results from the three tests coincided with each other.Conclusion The positive expression level of TNF-α is related to the severity degree of MP infection.

Key words: Mycoplasma pneumoniae, tumor necrosis factor-&alpha, euzymelinked immunosorbent assay, immunohistochemical technology

中图分类号:

R563.15

宋星宇，车广华，高航，胡起波，许忠，潘薇，徐坤 . 肿瘤坏死因子α在肺炎支原体感染小鼠主要脏器和血清中的表达及其相关机制[J]. 吉林大学学报(医学版), 2013, 39(6): 1195-1200.

SONG Xing-yu，CHE Guang-hua，GAO Hang，HU Qi-bo，XU Zhong，PAN Wei，XU Kun . Expressions of tumor necrosis factor-α in main organs and  serum of mice infected by Mycoplasma pneumoniae and their mechanisms[J]. Journal of Jilin University Medicine Edition, 2013, 39(6): 1195-1200.

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肿瘤坏死因子α在肺炎支原体感染小鼠主要脏器和血清中的表达及其相关机制

Expressions of tumor necrosis factor-α in main organs and  serum of mice infected by Mycoplasma pneumoniae and their mechanisms

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